Electrochemical/electrochemiluminescence sensors based on vertically-ordered mesoporous silica films for biomedical analytical applications.

Vertically-ordered mesoporous silica films (VMSF, also named as silica isoporous membranes) have shown tremendous potential in the field of electroanalytical sensors due to their unique features in terms of controllable and ultrasmall nanopores, high molecular selectivity and permeability, and mechanical stability. This review will present the recent progress on the biomedical analytical applications of VMSF, focusing on the small biomolecules, diseases-related biomarkers, drugs and cancer cells. Finally, conclusions with recent developments and future perspective of VMSF in the relevant fields will be envisioned.

Silica Nanochannels as Nanoreactors for the Confined Synthesis of Ag NPs to Boost Electrochemical Stripping Chemiluminescence of the Luminol-O2 System for the Sensitive Aptasensor.

Herein, we, for the first time, synthesize silver nanoparticles (Ag NPs) within the nanochannels of amino group-functionalized vertically ordered mesoporous silica films (NH2-VMSF) and investigate their coreaction accelerator role in the luminol-dissolved oxygen (O2) electrochemical stripping chemiluminescence (ESCL) system. The synthesized Ag NPs are capable of electrocatalytic reduction of O2 to superoxide radicals, and meanwhile, sliver ions (Ag+) electrochemically stripped from Ag NPs can promote the amount of luminol anion radicals, generating the boosted ECL intensity of the luminol-dissolved O2 system. This proposed Ag NPs@NH2-VMSF on the indium tin oxide electrode was applied to construct the ESCL aptasensor for quantitative determination of prostate-specific antigen (PSA), yielding a low detection limit [0.19 pg/mL (S/N = 3)] and a broad linear dynamic range (1 pg/mL to 100 ng/mL). Furthermore, good analytical performance of PSA in serum with satisfactory recoveries and low relative standard deviation values is achieved by our developed ESCL aptasensor, rendering it a convenient and sensitive method for PSA determination in clinical applications and further broadening the strategy of ESCL techniques.

Nanogold amplified electrochemiluminescence/electrochemistry in bipolar silica nanochannel array for ultrasensitive detection of SARS-CoV-2 pseudoviruses.

The prompt and accurate point-of-care test (POCT) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in infected persons or virus-containing environmental samples is of great importance. The present work reports a highly integrated electrochemiluminescence/electrochemical (ECL/EC) sensor for determination of SARS-CoV-2 pseudoviruses, in which bio-recognition element (SARS-CoV-2 IgG antibody), bifunctional probe (tris (2,2'-bipyridyl) ruthenium (Ru(bpy)32+)), and amplification material (gold nanoparticles (Au NPs)) are designed into bipolar silica nanochannel array (bp-SNA). bp-SNA consisting of homogeneous two-layer mesoporous silica films bears inner silanol groups and outer amino groups, generating a solid "electrostatic nanocage" for stable confinement of Ru(bpy)32+ and Au NPs inside the nanochannels and further providing functional sites for covalent modification of SARS-CoV-2 IgG antibody. Owing to the preconcentration capacity of bp-SNA and amplified effect of Au NPs, ECL or EC signals of Ru(bpy)32+ can be remarkably promoted and thereby increase the analytical performance, which can be diminished by immunorecognization of target SARS-CoV-2 pseudoviruses on the sensing interface. The developed integrated ECL/EC sensor based on Ru@AuNPs/bp-SNA modified solid indium tin oxide electrode enables the sensitive analysis of SARS-CoV-2 pseudoviruses by ECL mode with a linear range of 50 TU mL-1-5000 TU mL-1, as well as the EC mode with a linear range of 100 TU mL-1-5000 TU mL-1. Moreover, the designed sensor showed satisfactory results in the analyses of saliva and pond water samples. When flexible electrode substate (polyethylene terephthalate) is employed, Ru@AuNPs/bp-SNA has great potential to integrate with KN95 face masks for direct detection of SARS-CoV-2 pseudoviruses produced from breathing, talking and coughing processes, which could provide an efficient platform for POCT diagnosis.

Rational Design of an In-Situ Polymer-Inorganic Hybrid Solid Electrolyte Interphase for Realising Stable Zn Metal Anode under Harsh Conditions.

The in-depth understanding of the composition-property-performance relationship of solid electrolyte interphase (SEI) is the basis of developing a reliable SEI to stablize the Zn anode-electrolyte interface, but it remains unclear in rechargeable aqueous zinc ion batteries. Herein, a well-designed electrolyte based on 2 M Zn(CF3SO3)2-0.2 M acrylamide-0.2 M ZnSO4 is proposed. A robust polymer (polyacrylamide)-inorganic (Zn4SO4(OH)6.xH2O) hybrid SEI is in situ constructed on Zn anodes through controllable polymerization of acrylamide and coprecipitation of SO4 2- with Zn2+ and OH-. For the first time, the underlying SEI composition-property-performance relationship is systematically investigated and correlated. The results showed that the polymer-inorganic hybrid SEI, which integrates the high modulus of the inorganic component with the high toughness of the polymer ingredient, can realize high reversibility and long-term interfacial stability, even under ultrahigh areal current density and capacity (30 mA cm-2~30 mAh cm-2). The resultant Zn||NH4V4O10 cell also exhibits excellent cycling stability. This work will provide a guidance for the rational design of SEI layers in rechargeable aqueous zinc ion batteries.

Spectroscopic studies of the role of dissolved organic matter in acenaphthene photodegradation in liquid water and ice.

The effect of concentration and origin of dissolved organic matter (DOM) on acenaphthene (Ace) photodegradation in liquid water and ice was investigated, and the components in DOM which were involved in Ace photodegradation were identified. The DOM samples included Suwannee River fulvic acid (SRFA), Elliott soil humic acid (ESHA), and an effluent organic matter (EfOM) sample. Due to the production of hydroxyl radical (•OH) and triplet excited-state DOM (3DOM*) which react with Ace, DOM had promotion effects on Ace photodegradation. However, the promotion effects of DOM were prevailed over by their suppressing effect of DOM including screening light effect, intermediates reducing effect and RS quenching effect, and thus, the photodegradation rates of Ace decreased in the presence of the three DOM with concentrations of 0.5-7.5 mg C/L in liquid water and ice. ESHA had higher light absorption and thus had higher screening light effect on Ace photodegradation in liquid water than SRFA and EfOM. At each DOM concentration, ESHA exhibited higher promotion effect on Ace photodegradation than SRFA and EfOM, in liquid water and ice. The binding of Ace with DOM was indicated by decreases in fluorescence intensity of Ace when coexisted with DOM. However, the binding of Ace to DOM played an unimportant role in suppressing Ace photodegradation. The photodegradation behavior of fluorophores in Ace with DOM present in ice was not similar to that in liquid water. C-O, C═O, carboxyl groups O-H and aliphatic C-H functional groups in DOM were involved in the interaction of DOM with Ace. The presence of Ace seemed to have no influence on the photodegradation behavior of functional groups in DOM.

Solid Electrochemiluminescence Sensor by Immobilization of Emitter Ruthenium(II)tris(bipyridine) in Bipolar Silica Nanochannel Film for Sensitive Detection of Oxalate in Serum and Urine.

Convenient and highly sensitive detection of oxalate ions in body fluids is of crucial significance for disease prevention, diagnosis, and monitoring of treatment effectiveness. Establishing a simple solid-state electrochemiluminescence (ECL) sensing system for highly sensitive detection of oxalate ions is highly desirable. In this work, a solid ECL sensor was fabricated by immobilizing the commonly used emitter ruthenium(II)tris(bipyridine) (Ru(bpy)32+) on a double-layered bipolar silica nanochannel array film (bp-SNA)-modified electrode, enabling sensitive detection of oxalate ions in serum or urine samples. Cost-effective and readily available indium tin oxide (ITO) was used as the supporting electrode. Convenient fabrication of multiple negatively charged SNA (n-SNA)-modified ITO electrodes was achieved through the one-step Stöber solution growth method. Subsequently, a positive outer layer film (p-SNA) was rapidly prepared using an electrochemical-assisted self-assembly method. The double-layered bipolar silica nanochannel array film achieved stable immobilization of Ru(bpy)32+ on the electrode surface, facilitated by the electrostatic adsorption of Ru(bpy)32+ by n-SNA and the electrostatic repulsion by p-SNA. Utilizing oxalate ions as a co-reactant for Ru(bpy)32+, combined with the electrostatic enrichment of oxalate ions by p-SNA, the constructed sensor enabled highly sensitive detection of oxalate ions ranging from 1 nM to 25 µM and from 25 µM to 1 mM, with a detection limit (LOD) of 0.8 nM. The fabricated ECL sensor exhibited high selectivity and good stability, making it suitable for ECL detection of oxalate ions in serum and urine samples.

Effect of carboxyl and hydroxyl groups attached to the benzene ring on the photodegradation of polycyclic aromatic hydrocarbons in ice.

The effect of carboxyl and hydroxyl groups attached to the benzene ring on the photodegradation of anthracene (Ant) and pyrene (Pyr) in ice was investigated. The present study aims to explore the inhibition mechanism of five dissolved organic matter (DOM) model compounds' materials such as benzoic acid, o-hydroxybenzoic acid, m-hydroxybenzoic acid, p-hydroxybenzoic acid, and 3-phenyl propionic acid on the degradation of Ant and Pyr in ice. The photodegradation rate of Ant and Pyr were 50.33 and 37.44% in ice, with the photodegradation rate of Ant being greater than that of Pyr. The five DOM model compounds inhibited the photolysis of Ant and Pyr, and the influence mechanism on the photodegradation of Ant and Pyr depended upon the types and positions of functional groups on the benzene. Among them, the structure in which the carboxyl group was directly connected to the benzene ring and carboxyl was located at the ortho position of a hydroxy group had a strong inhibitory effect on the photodegradation of Ant and Pyr. Light-screening effects and quenching effects were the main inhibiting mechanism, and the binding ability of DOM model compounds material and PAHs is dominantly correlated with its inhibiting effect.

Electrochemiluminescence Aptasensor with Dual Signal Amplification by Silica Nanochannel-Based Confinement Effect on Nanocatalyst and Efficient Emitter Enrichment for Highly Sensitive Detection of C-Reactive Protein.

The rapid and sensitive detection of the important biomarker C-reactive protein (CRP) is of great significance for monitoring inflammation and tissue damage. In this work, an electrochemiluminescence (ECL) aptasensor was fabricated based on dual signal amplification for the sensitive detection of CRP in serum samples. The sensor was constructed by modifying a silica nanochannel array film (SNF) on a cost-effective indium tin oxide (ITO) electrode using the Stöber solution growth method. Gold nanoparticles (AuNPs) were grown in situ within the nanochannels using a simple electrodeposition method as a nanocatalyst to enhance the active electrode area as well as the ECL signal. The negatively charged nanochannels also significantly enriched the positively charged ECL emitters, further amplifying the signal. The recognition aptamer was covalently immobilized on the outer surface of SNF after modification with epoxy groups, constructing the aptasensor. In the presence of CRP, the formation of complexes on the recognitive interface led to a decrease in the diffusion of ECL emitters and co-reactants to the supporting electrode, resulting in a reduction in the ECL signal. Based on this mechanism, ECL detection of CRP was achieved with a linear range of 10 pg/mL to 1 µg/mL and a low limit of detection (7.4 pg/mL). The ECL aptasensor developed in this study offers advantages such as simple fabrication and high sensitivity, making promising applications in biomarker detection.

Label-Free Homogeneous Electrochemical Aptasensor Based on Size Exclusion/Charge-Selective Permeability of Nanochannel Arrays and 2D Nanorecognitive Probe for Sensitive Detection of Alpha-Fetoprotein.

The labeling-free and immobilization-free homogeneous aptamer sensor offers advantages including simple operation, low cost, and high sensitivity, demonstrating great potential in rapid detection of tumor biomarkers in biological samples. In this work, a labeling-free and immobilization-free homogeneous aptamer sensor was conveniently fabricated by combining size exclusion and charge-selective penetration of a nanochannel-modified electrode and two-dimensional (2D) nanorecognition probe which can realize selective and highly sensitive detection of alpha-fetoprotein (AFP) in serum. Vertically ordered mesoporous silica film (VMSF) with ultra-small, uniform, and vertically aligned nanochannels was easily grown on the simple, low-cost, and disposable indium tin oxide (ITO) electrode. Through π-π interaction and electrostatic force, the AFP aptamer (Apt) and electrochemical probe, tris(bipyridine)ruthenium(II) (Ru(bpy)32+), were coloaded onto graphene oxide (GO) through simple incubation, forming a 2D nanoscale recognition probe (Ru(bpy)32+/Apt@GO). Owing to the size exclusion effect of VMSF towards the 2D nanoscale probe, the electrochemical signal of Ru(bpy)32+/Apt@GO could not be detected. In the presence of AFP, the specific binding of AFP to the aptamer causes the dissociation of the aptamer and Ru(bpy)32+ from GO, resulting in their presence in the solution. The efficient electrostatic enrichment towards Ru(bpy)32+ by negatively charged VMSF allows for high electrochemical signals of free Ru(bpy)32+ in the solution. Linear determination of AFP ranged from 1 pg/mL to 1000 ng/mL and could be obtained with a low limit of detection (LOD, 0.8 pg/mL). The high specificity of the adapter endowed the constructed sensor with high selectivity. The fabricated probe can be applied in direct determination of AFP in serum.

Effect of dissolved organic matter on sulfachloropyridazine photolysis in liquid water and ice.

Dissolved organic matter (DOM) is an ubiquitous component of environmental snow and ice, which can absorb light and produce reactive species (RS) and thus is of importance in ice photochemistry. The photodegradation of sulfachloropyridazine (SCP) without and with DOM present in liquid water and ice were investigated in this study. The photodegradation rate constants for SCP without DOM present was enhanced by 52.5 % in ice relative to liquid water, likely due to the enhanced role of SCP self-sensitized RS in ice. DOM significantly promoted SCP photolysis in both liquid water and ice, which was mainly attributed to roles of singlet oxygen (1O2) and triplet excited-state DOM (3DOM*) generated from DOM. 1O2 production from DOM was significantly enhanced in ice relative to liquid water. Hydroxyl radical (•OH) production from DOM in ice was similar to those in liquid water. Enhancement in 3DOM* production in ice was observed at low DOM concentrations. Suwannee River Fulvic Acid (SRFA) and Elliott Soil Humic Acid (ESHA) exhibited differences in RS production in liquid water and ice, as well as in enhancement of 1O2 and 3DOM* produced in ice relative to liquid water. DOM induced reaction pathways of SCP different from those without DOM present, and therefore affected toxicity of SCP photoproducts. There were differences in photodegradation pathways of SCP as well as in toxicity of SCP photoproducts between liquid water and ice.

Preparation and characterization of a homogeneous immunoassay for point-of-care testing (POCT) of procalcitonin (PCT).

Procalcitonin (PCT) has been recognized as a specific and early marker for microbial infection and sepsis. Sensitive measuring interaction-triggered luminescence experiment (SMILE), a homogeneous immunoassay method, was established for point-of-care testing (POCT) of PCT. SMILE is achieved through the principle of double antibody sandwich, where two antibodies immobilized on the surface of polystyrene microspheres (donor and acceptor beads) bind to the PCT antigen. The donor bead contains phthalocyanine dye (luminol chemiluminescent substance) and the acceptor bead contains dimethylthiophene derivatives and Eu chelates. Therefore, singlet oxygen can be transferred when the distance between donor and acceptor beads is within 200 nm, generating detectable luminescent signals. Scanning electron microscopy (SEM) was used to detect the diameter and polymer dispersity index (PDI) of microspheres before and after binding with antibodies to characterize the immobilization of antibodies. The reaction conditions for antibody immobilization including pH, mass ratio and reaction time have also been optimized. The limit of quantitation (LOQ) of the SMILE method (0.01 ng mL-1) was lower than that of the LFI method (0.1 ng mL-1), the working range (0.01-500 ng mL-1) was wider than that of the LFI method (0.1-50 ng mL-1), and the assay time (10 min) was shorter than that of the LFI method (15 min). So, SMILE is more suitable for POCT of PCT compared with lateral flow immunochromatography (LFI), which is the most used measuring method, due to its advantages of simple operation, saving time, convenience, wide detection range, and high sensitivity and accuracy.

Facile Synthesis of Iron and Nitrogen Co-Doped Carbon Dot Nanozyme as Highly Efficient Peroxidase Mimics for Visualized Detection of Metabolites.

Visual detection based on nanozymes has great potential for the rapid detection of metabolites in clinical analysis or home-based health management. In this work, iron and nitrogen co-doped carbon dots (Fe,N-CDs) were conveniently synthesized as a nanozyme for the visual detection of glucose (Glu) or cholesterol (Chol). Using inexpensive and readily available precursors, Fe,N-CDs with peroxidase-like activity were conveniently prepared through a simple hydrothermal method. Co-doping of Fe and N atoms enhanced the catalytic activity of the nanozyme. The nanozyme had a low Michaelis constant (Km) of 0.23 mM when hydrogen peroxide (H2O2) was used as the substrate. Free radical trapping experiments revealed that the reactive oxygen species (ROS) generated in the nanozyme-catalyzed process were superoxide anion radicals (•O2-), which can oxidize colorless 3,3',5,5'-tetramethylbenzidine (TMB) to generate blue oxidation product (ox-TMB) with characteristics absorbance at 652 nm. Based on this mechanism, a colorimetric sensor was constructed to detect H2O2 ranging from 0.1 µM to 200 µM with a detection limit (DL) of 75 nM. In the presence of glucose oxidase (Gox) or Chol oxidase (Chox), Glu or Chol was oxidized, respectively, and generated H2O2. Based on this, indirect detection of Glu and Chol was realized with linear detection ranges of 5-160 µM and 2-200 µM and DLs of 2.8 µM and 0.8 µM, respectively. A paper-based visual detection platform was fabricated using Fe,N-CDs as nanozyme ink to prepare testing paper by inkjet printing. Using a smartphone to record the RGB values of the testing paper after the reaction, visual detection of Glu and Chol can be achieved with linear detection ranges of 5-160 µM (DL of 3.3 µM) and 2-200 µM (DL of 1.0 µM), respectively.

Homogeneous Electrochemical Aptamer Sensor Based on Two-Dimensional Nanocomposite Probe and Nanochannel Modified Electrode for Sensitive Detection of Carcinoembryonic Antigen.

A rapid and convenient homogeneous aptamer sensor with high sensitivity is highly desirable for the electrochemical detection of tumor biomarkers. In this work, a homogeneous electrochemical aptamer sensor is demonstrated based on a two-dimensional (2D) nanocomposite probe and nanochannel modified electrode, which can realize sensitive detection of carcinoembryonic antigen (CEA). Using π-π stacking and electrostatic interaction, CEA aptamer (Apt) and cationic redox probe (hexaammineruthenium(III), Ru(NH3)63+) are co-loaded on graphite oxide (GO), leading to a 2D nanocomposite probe (Ru(NH3)63+/Apt@GO). Vertically ordered mesoporous silica-nanochannel film (VMSF) is easily grown on the supporting indium tin oxide (ITO) electrode (VMSF/ITO) using the electrochemical assisted self-assembly (EASA) method within 10 s. The ultrasmall nanochannels of VMSF exhibits electrostatic enrichment towards Ru(NH3)63+ and size exclusion towards 2D material. When CEA is added in the Ru(NH3)63+/Apt@GO solution, DNA aptamer recognizes and binds to CEA and Ru(NH3)63+ releases to the solution, which can be enriched and detected by VMSF/ITO electrodes. Based on this mechanism, CEA can be an electrochemical detection ranging from 60 fg/mL to 100 ng/mL with a limit of detection (LOD) of 14 fg/mL. Detection of CEA in human serum is also realized. The constructed homogeneous detection system does not require the fixation of a recognitive aptamer on the electrode surface or magnetic separation before detection, demonstrating potential applications in rapid, convenient and sensitive electrochemical sensing of tumor biomarkers.

Label-Free Electrochemical Aptasensor Based on the Vertically-Aligned Mesoporous Silica Films for Determination of Aflatoxin B1.

Herein we report a highly specific electrochemical aptasenseor for AFB1 determination based on AFB1-controlled diffusion of redox probe (Ru(NH3)63+) through nanochannels of AFB1-specific aptamer functionalized VMSF. A high density of silanol groups on the inner surface confers VMSF with cationic permselectivity, enabling electrostatic preconcentration of Ru(NH3)63+ and producing amplified electrochemical signals. Upon the addition of AFB1, the specific interaction between the aptamer and AFB1 occurs and generates steric hindrance effect on the access of Ru(NH3)63+, finally resulting in the reduced electrochemical responses and allowing the quantitative determination of AFB1. The proposed electrochemical aptasensor shows excellent detection performance in the range of 3 pg/mL to 3 µg/mL with a low detection limit of 2.3 pg/mL for AFB1 detection. Practical analysis of AFB1 in peanut and corn samples is also accomplished with satisfactory results by our fabricated electrochemical aptasensor.

Reagentless Electrochemical Detection of Tumor Biomarker Based on Stable Confinement of Electrochemical Probe in Bipolar Silica Nanochannel Film.

The development of simple and probe-integrated aptamer sensors for the electrochemical detection of tumor biomarkers is of great significance for the diagnosis of tumors and evaluation of prognosis. In this work, a probe-integrated aptamer sensor is demonstrated based on the stable confinement of an electrochemical probe in a bipolar nanochannel film, which can realize the reagentless electrochemical detection of the tumor biomarker carcinoembryonic antigen (CEA). To realize the stable immobilization of a large amount of the cationic electrochemical probe methylene blue (MB), a two-layer silica nanochannel array (SNF) with asymmetric charge was grown on the supporting electrode from bipolar SNF (bp-SNF). The inner SNF is negatively charged (n-SNF), and the outer-layer SNF is positively charged (p-SNF). The dual electrostatic interaction including the electrostatic adsorption from n-SNF and the electrostatic repulsion from p-SNF achieve the stable confinement of MB in bp-SNF. The recognitive interface is fabricated by the covalent immobilization of the CEA aptamer on the outer surface of bp-SNF, followed by the blocking of non-specific binding sites. Owing to the stable and abundant immobilized probes and highly specific aptamer interface, the developed aptamer sensor enables the sensitive detection of CEA in the range of 1 pg/mL to 1 µg/mL with a low limit of detection (LOD, 0.22 pg/mL, S/N = 3). Owing to the high selectivity and stability of the developed biosensor, reagentless electrochemical detection of CEA in serum was realized.

Photoredox Microfluidic Synthesis of Trifluoromethylated Amino Acids.

Fluorinated amino acids are a class of highly valuable building blocks that are widely employed in biological science and pharmaceutical industry for improved stability, activity, and folding property of proteins. However, the synthetic approach has conventionally been constrained by harsh conditions and limited substrate range. We demonstrate a general synthetic protocol for photoinduced α-CF3 amino acids using continuous flow technology that benefits from enhanced fusion and precise control of reaction time, making it potentially useful in large-scale peptide synthesis.

Dual-Mode Sensing Platform for Cancer Antigen 15-3 Determination Based on a Silica Nanochannel Array Using Electrochemiluminescence and Electrochemistry.

An electrochemiluminescence-electrochemistry (ECL-EC) dual-mode sensing platform based on a vertically-ordered mesoporous silica films (VMSF) modified electrode was designed here for the sensitive and selective determination of cancer antigen 15-3 (CA 15-3), a specific biomarker of breast cancer. VMSF was assembled through a rapid electrochemically assisted self-assembly (EASA) method and plays a crucial role in signal amplification via a strong electrostatic interaction with the positively charged bifunctional probe Ru(bpy)32+. To construct the biorecognition interface, epoxy functional silane was linked to the surface of VMSF for further covalent immobilization of the antibody. As a benefit from the specific combination of antigen and antibody, a non-conductive immunocomplex layer was formed in the presence of CA 15-3, leading to the hinderance of the mass and electron transfer of the probes. Based on this strategy, the dual-mode determination of CA 15-3 ranging from 0.1 mU/mL to 100 mU/mL with a LOD of 9 µU/mL for ECL mode, and 10 mU/mL to 200 U/mL with a LOD of 5.4 mU/mL for EC mode, was achieved. The proposed immunosensor was successfully employed for the detection of CA 15-3 in human serum without tedious pretreatment.

A DMD case caused by X chromosome rearrangement.

Duchenne/Becker muscular dystrophy (DMD/BMD) is one of the most common progressive muscular dystrophy diseases with X-linked recessive inheritance. It is mainly caused by the deletion, duplication and point mutation of DMD gene. In rare cases, it is also caused by the destruction of DMD gene by chromosomal structural rearrangement. Here, we report a case of Duchenne/Becker Muscular dystrophy (DMD/BMD) with typical symptoms but unknown genetic defects after MLPA and next generation sequencing tests in other hospitals. Interestingly, we find a pericentric inversion of X chromosome (Chr.X: g. [31939463-31939465del; 31939466-131765063 inv; 131765064-131765067del]) in this patient. We then use the karyotyping, FISH, long-read sequencing and Sanger sequencing technologies to characterize the chromosome rearrangement. We find that this chromosomal aberration disrupt both the DMD gene and the HS6ST2 gene. The patient present with typical DMD symptoms such as muscle weakness, but no obvious symptoms of Paganini-Miozzo syndrome. Our results suggest that the destruction of DMD gene by structural rearrangement is also one of the important causes of DMD. Therefore, we suggest to provide further genetic testing for those DMD patients with unknown genetic defects through routine genetic testing. Cost-effective karyotyping and FISH should be considered firstly to identify chromosome rearrangements. Long-read sequencing followed by Sanger sequencing could be useful to locate the precise breakpoints. The genetic diagnosis of this case made it possible for reproductive intervention in the patient's family.

Equipment of Vertically-Ordered Mesoporous Silica Film on Electrochemically Pretreated Three-Dimensional Graphene Electrodes for Sensitive Detection of Methidazine in Urine.

Direct, rapid, and sensitive detection of drugs in complex biological samples is essential for drug abuse control and health risk assessment. In this work, an electrochemical sensor was fabricated based on equipment of vertically-ordered mesoporous silica film (VMSF) on an electrochemically pre-treated three-dimensional graphene electrode (p-3DG), which can achieve direct and sensitive determination of methylthiopyridazine (TR) in urine. Three-dimensional graphene (3DG) with a continuous and interpenetrating graphene network was used as the supporting electrode and simple electrochemical polarization was employed to pre-treat 3DG to improve surface hydrophilicity and electrocatalytic performance. VMSF was easily grown using an electrochemical assisted self-assembly method within 10 s and was stably bound to the p-3DG surface. The nanochannel array on the as-prepared VMSF/p-3DG sensor enriched positively charged TR, leading to significantly improved electrochemical signal. Combined with the high electric activity of p-3DG and the enrichment of nanochannels, VMSF/p-3DG realized sensitive determination of TR ranging from 50 nM to 10 µM with a low detection limit (DL, 30 nM). Owing to the anti-fouling and anti-interference performance of VMSF, the common electroactive molecules including ascorbic acid (AA) and uric acid (UA) did not interfere with the detection. In addition, the detection of TR in buffer and urine exhibited similar sensitivity. Accurate detection of TR in urine was realized.

Associations of muscle and adipose tissue parameters with long-term outcomes in middle and low rectal cancer: a retrospective cohort study.

OBJECTIVE: To investigate the role of preoperative body composition analysis for muscle and adipose tissue distribution on long-term oncological outcomes in patients with middle and low rectal cancer (RC) who received curative intent surgery. METHODS: A total of 155 patients with middle and low rectal cancer who underwent curative intent surgery between January 2014 and December 2016 were included for the final analysis. Skeletal muscle area (SMA), skeletal muscle radiodensity (SMD), visceral fat area (VFA) and mesorectal fat area (MFA) were retrospectively measured using preoperative CT images. To standardize the area according to patient stature, SMA was divided by the square of the height (m2) and the skeletal muscle mass index (SMI, cm2/m2) was obtained. Each median values of the distribution in male and female served as cut-off point for SMI, SMD, VFA, and MFA, respectively. Univariate and multivariate analysis were performed to evaluate the association between body composition and long-term oncological outcomes. Overall survival (OS) measured in months from the day of primary surgery until death for any cause. Disease-free survival (DFS) was defined as the interval between surgery and tumor recurrence. The Kaplan-Meier method with log-rank testing was used to validate prognostic biomarkers. Intraclass correlation coefficient (ICC) was used to evaluate interobserver and intraobserver reproducibility for SMA, SMD, MFA,VFA. RESULTS: During the follow-up period, 42 (27.1%) patients had tumor recurrence; 21 (13.5%) patients died. The sex-specific median value of SMI was 28.6 cm2/m2 for females and 48.2 cm2/m2 for males. The sex-specific median value of SMD was 34.7 HU for females and 37.4 HU for males. The sex-specific median value of VFA was 123.1 cm2 for females and 123.2 cm2 for males. The sex-specific median value of MFA was 13.8 cm2 for females and 16.0 cm2 for males. In the Cox regression multivariate analysis, SMI (P = 0.036), SMD (P = 0.022), and postoperative complications grades (P = 0.042) were significantly different between death group and non-death group; SMD (P = 0.011) and MFA (P = 0.022) were significantly different between recurrence group and non-recurrence group. VFA did not show any significant differences. By the Kaplan-Meier method with log-rank testing, DFS was significantly longer in patients with high-MFA (P = 0.028) and shorter in patients with low-SMD (P = 0.010), OS was significantly shorter in patients with low-SMI (P = 0.034) and low-SMD (P = 0.029). CONCLUSIONS: Quantitative evaluation of skeletal muscle mass and adipose tissue distributions at initial diagnosis were important predictors for long-term oncologic outcomes in RC patients. SMD and SMI were independent factors for predicting OS in patients with middle and low rectal cancer who had radical surgery. SMD and MFA were independent factors for predicting DFS in patients with middle and low rectal cancer who had radical surgery.